Intermediates for preparing 7-acyl-3-(sulfonic acid and sulfamoyl substituted tetrazolylthiomethyl)cephalosporins

ABSTRACT

The compounds of this invention are cephalosporins having various acyl substituents at the 7-position and a sulfonic acid or sulfamoyl substituted tetrazolyl thiomethyl group at the 3-position of the cephem nucleus and intermediates for the preparation thereof. The 7-acylated compounds have antibacterial activity.

This is a division of application Ser. No. 687,792 filed May 19, 1976,now U.S. Pat. No. 4,048,311, which is a continuation-in-part ofapplication Ser. No. 647,394 filed Jan. 8, 1976, now abandoned, which isa continuation-in-part of application Ser. No. 559,609 filed Mar. 18,1975, now abandoned.

This invention comprises a new series of cephalosporin compounds whichhave antibacterial activity when administered parenterally and tointermediates for the preparation thereof. In particular, the structuresof the biologically active cephalosporin compounds of this invention arecharacterized by having a sulfonic acid or sulfamoyl substitutedtetrazolyl thiomethyl group at the 3-position of the cephem nucleus.Also, this invention extends to methods and compositions for treatingcertain bacterial infections using these new compounds as well as tocertain chemical intermediates and methods for preparing the compoundsdescribed hereafter.

The compounds of this invention are represented by the followingstructural formula: ##STR1## in which: EACH INDIVIDUAL R¹ is hydrogen orlower alkyl;

N is one to ten;

R² is hydroxy, amino, lower alkylamino or di(lower)alkylamino; and

R³ is an acyl group selected from the group consisting of: ##STR2##where: X is thienyl, dihydrophenyl; phenyl; phenyl mono-substituted withhydroxy, hydroxymethyl, formamido, ureido or carboxymethylamino; or3-fluoro-4-hydroxyphenyl;

A is NH₂, OH, COOH or SO₃ H; or formyloxy when X is phenyl;

Y is thienyl, tetrazolyl, cyano, sydnone or aminomethylphenyl;

Z is methyl, trifluoromethyl, trifluoroethyl, cyanomethyl or pyridyl;and

M is zero to two,

Or a non-toxic pharmaceutically acceptable salt thereof.

As used herein, the term "lower alkyl" refers to groups having from oneto four carbon atoms, preferably methyl and ethyl.

It will be recognized that the 4-carboxylic acid group of the compoundsof Formula I may be readily esterified by methods well known to the art.These esters include, for example, simple alkyl and aryl esters as wellas esters which are easily cleaved, within the body, to the parent acidsuch as indanyl, pivaloyloxymethyl, acetoxymethyl, propionyloxymethyl,glycyloxymethyl, phenylglycyloxymethyl and thienylglycyloxymethyl estersand others. Of course, when A is COOH, this group may be similarlyesterified. All such esters are included within the scope of thisinvention.

A selected group of compounds of this invention are represented byFormula I where R¹ is hydrogen; n is one to five; R² is hydroxy, amino,lower alkylamino or di(lower)alkylamino: ##STR3## X is thienyl,dihydrophenyl, phenyl, phenyl mono-substituted with hydroxy,hydroxymethyl, formamido, ureido or carboxymethylamino, or3-fluoro-4-hydroxyphenyl; A is NH₂, OH, COOH or SO₃ H; Y is thienyl,tetrazolyl, cyano, sydnone or aminomethylphenyl; Z is methyl,trifluoromethyl, trifluoroethyl, cyanomethyl or pyridyl and m is zero totwo.

Another group of compounds of this invention are represented by FormulaI where R¹ is hydrogen; n is one to five; R² is hydroxy, amino, loweralkylamino or di(lower)alkylamino; X is phenyl or hydroxyphenyl; A isNH₂ or OH; Y is thienyl; Z is methyl, trifluoromethyl or trifluoroethyland m is zero to two.

Yet another group are those compounds represented by Formula I where R¹is hydrogen; n is one to five; R² is hydroxy or amino; X is phenyl or4-hydroxyphenyl; A is NH₂ or OH; Y is thienyl; Z is trifluoromethyl andm is zero.

Examples of representative 7-acyl substituents (R³ NH--) of thecompounds of Formula I are listed below:

α-hydroxyphenylacetamido

α-aminophenylacetamido

α-amino-4-hydroxyphenylacetamido

trifluoromethylthioacetamido

methylthioacetamido

2,2,2-trifluoroethylsulfinylacetamido

thienylacetamido

tetrazolylacetamido

cyanoacetamido

α-carboxythienylacetamido

α-carboxyphenylacetamido

α-sulfophenylacetamido

methylsulfonylacetamido

cyanomethylthioacetamido

α-amino-4-carboxymethylaminophenylacetamido

α-amino-3-fluoro-4-hydroxyphenylacetamido

3-sydnoneacetamido

4-pyridylthioacetamido

2-aminomethylphenylacetamido.

Representative substituted tetrazolyl groups are the following:

1-sulfomethyltetrazolyl

1-(2-sulfoethyl)tetrazolyl

1-(2-sulfamoylethyl)tetrazolyl

1-(3-sulfopropyl)tetrazolyl

1-(3-sulfamoylpropyl)tetrazolyl

1-(5-sulfopentyl)tetrazolyl

1-(5-sulfamoylpentyl)tetrazolyl.

Some examples of the compounds of this invention are7-D-mandelamido-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid,7-D-mandelamido-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid,7-D-mandelamido-3-[1-(5-sulfopentyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid,7-D-mandelamido-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid,7-(2-thienylacetamido-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid,7-(2-thienylacetamido)-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid,7-trifluoromethylthioacetamido-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid, 7-(2-thienylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid and7-trifluoromethylthioacetamido-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid.

Cephalosporin derivatives having 7-acyl substituents as defined aboveare all documented in the prior art. Substitution by a substitutedS-heterocyclicthiomethyl group (--CH₂ SHet) at the 3-position of thecephem nucleus is also known and is disclosed in Netherlands Patent6916151 where Het is, among others, tetrazolyl substituted with, interalia, carboxy, carbalkoxy, alkoxyalkylaminocarbonyl anddialkylaminoalkylaminocarbonyl and in Japanese patent 7205550 where Hetincludes tetrazolyl substituted with --(CH₂)_(n) R³ where n is 0 to 3and R³ includes alkoxycarbonyl, carboxy, N-alkoxyalkylcarbamoyl anddialkylamino. Recently issued U.S. Pat. No. 3,819,623 disclosescephalosporins bearing a 7-heterocyclicacetamido or7-heterocyclicthioalkylacetamido group and having in the 3-position,inter alia, thiomethyltetrazolyl substituted with carboxy,alkoxycarbonyl, carboxyalkyl, alkoxycarbonylalkyl anddialkylaminoalkylaminocarbonylalkyl. No compounds containing applicant's3-(sulfonic acid or sulfamoyl substituted tetrazolyl)thiomethyl moietydisclosed herein are believed to be known to the art.

The compounds of Formula I are prepared by acylating7-aminocephalosporanic acid with an appropriately protected acylatingagent and then displacing the 3-acetoxy group with the desiredsubstituted tetrazole thiol with subsequent removal of the protectivegroup(s). The substituted tetrazole thiols of the formula: ##STR4## inwhich: each individual R¹ is hydrogen or lower alkyl;

n is one to ten; and

R² is hydroxy, amino, lower alkylamino or di(lower)alkylamino,

are also objects of this invention, being important intermediates forproducing pharmaceutical end products as described herein.

The carboxylic acid group of the acylating agent is activated by any ofthe standard methods such as conversion to the mixed anhydride, acidchloride, acid imidazolide or activated ester. In addition, a reagentsuch as dicyclohexylcarbodiimide can be used provided that the carboxylgroup on the cephem nucleus is protected with an easily removableprotecting group such as a benzhydryl, t-butyl, trichloroethyl, benzyl,benzyloxymethyl, p-nitrophenyl, p-methoxyphenyl, p-methoxybenzyl orp-nitrobenzyl ester. When A is NH₂, the α-amino group of the acylatingagent is, preferably protected prior to acylation with an easilyremovable protective group known in the art such as t-butoxycarbonyl,trichloroethoxycarbonyl, benzyloxycarbonyl, the methyl acetoacetateadduct or similar groups commonly used in the synthesis of peptides.

Alternatively, the compounds of Formula I are prepared by acylation ofan appropriate 7-amino-3- -substituted tetrazolylthiomethylcephalosporin nucleus of Formula III: ##STR5## in which: each individualR¹ is hydrogen or lower alkyl;

n is one to ten;

R² is hydroxy, amino, lower alkylamino or di(lower)alkylamino; and

R⁴ is hydrogen or a protecting ester group, with an appropriateacylating agent followed by removal of the protective groups whenpresent.

The compounds of Formula III above are also considered as objects ofthis invention.

The protective groups can be removed according to methods well known tothe art, such as with trifluoroacetic acid when t-butyl ort-butoxycarbonyl protective groups are used. The resulting salt isconverted to the zwitterionic product or to the free acid by means of abasic ion exchange resin such as polystyrene-amine ion exchange resin(Amberlite IR-45) or else by basification of an aqueous solution of thesalt.

The acylating agents used as starting materials are either known orprepared by known methods.

The 7-amino-3-substituted tetrazolylthiomethylcephalosporin startingmaterials of Formula III are prepared from reaction of7-aminocephalosporanic acid or 7-formamidocephalosporanic acid, preparedby reaction of 7-aminocephalosporanic acid with formic acid and aceticanhydride, and a substituted tetrazole thiol of Formula II followed bytreatment with acid such as hydrochloric acid to remove the formyl groupwhen employed.

The substituted tetrazole thiols of Formula II where R² is hydroxy,lower alkylamino or di(lower)alkylamino are prepared by reaction of anN-alkyl dithiocarbamate, such as methyl 2-sulfoethyldithiocarbamate ormethyl 3-(N-t-butylsulfamoylpropyl)dithiocarbamate or its correspondingsodium or potassium salt with an azide such as sodium azide. The N-alkyldithiocarbamates are prepared by treatment of an aminoalkanesulfonicacid, for example 2-aminoethanesulfonic acid, or an amino (N-alkyl orN,N-dialkyl)sulfonamide such as 3-aminopropane-N-t-butyl-sulfonamide orits corresponding salt with carbon disulfide and an alkyl halide such asmethyl iodide in the presence of a base such as sodium or potassiumhydroxide.

The amino(N-alkyl or N,N-dialkyl)sulfonamides are prepared by reactionof an N-alkyl or N,N-dialkylphthalimidoalkylsulfonamide, obtained fromtreatment of a phthalimidoalkylsulfonyl halide, preferably chloride,with an alkyl- or dialkylamine and then with hydrazine. Thephthalimidoalkylsulfonyl halides are known or are prepared as describedby Winterbottom et al., J. Amer. Chem. Soc. 69:1393 (1947) and Griffinand Hey, J. Chem. Soc., 3334 (1952).

When R² is amino, the compounds of Formula II are prepared by removal ofthe N-alkyl group, which also serves as an amine protective group, fromthe corresponding N-alkylsulfamoylalkyltetrazole-5-thiol, preferably aN-t-butylsulfamoylalkyltetrazole-5-thiol, with, for example, anisole andtrifluoroacetic acid.

Certain compounds of this invention are capable of forming salts with,for example, the alkali metals such as sodium or potassium, the alkalineearth metals such as calcium or with the ammonium cation. When A is NH₂,the compounds can exist as the zwitterion or as either an acid or basesalt. These salts are prepared by standard methods using a wide varietyof non-toxic pharmaceutically acceptable acids and bases known in theart and are also considered as objects of this invention.

It will be recognized that due to the asymmetric α-carbon atom in the7-acetamido group of Formula I when ##STR6## and the potentiallyasymmetric carbon atom in the tetrazole sidechain, optical isomers willexist. Racemic or resolved products are obtained depending upon whethera racemic or resolved sidechain acid is used as an acylating agent andwhether a racemic or resolved tetrazole thiol is used. The resolvedsidechain acids are readily obtained from the racemic compounds byresolution according to well known methods, including fractionalcrystallization of a salt formed with an optically acid acid or base.All of the isomers, including separated isomers and mixtures thereof,are included within the scope of this invention.

The compounds of Formula I have exceptional antibacterial activityagainst both Gram-positive and Gram-negative organisms. Minimuminhibitory concentrations (MIC's) ranged from 0.2 to >200 μ g./ml. in invitro testing. These results are shown in Table 1 below forrepresentative compounds of Formula I. In vivo mouse protection data aregiven in Table 2. Compound names corresponding to numbers are given inTable 3.

    TABLE 1      MIC (μg./ml.) in vitro     Strep.        Entero. Serra. Entero.     Faecalis   Kleb. Kleb. Pseudo. Salmonella  aerog. marc. cloacae Proteus     Com- S. aureus S. aureus S. HH E. coli E. coli pneumo. pneumo. sp. ATCC     Shigella ATCC ATCC HH morgani pound HH 127 SK 23390 villaluz 34358 SK     12140 HH 33779 SK 4200 SK 1200 HH 63 12176 HH 117 13048 13880 31254     179    I 3.1, 0.8, 6.3, 50, 0.8, 3.1, 0.8, 0.2, >200, 0.8, 0.4, 3.1     12.5, 0.8, 3.1,  6 6 50 100 0.8 1.6 0.8 0.8 >200 0.4 0.4 13 13 3 3 II     1.6 0.8 6.3 50 0.8 1.6 0.8 0.4 >200 0.8 0.4 6.3 25 3.1 6.3 III 1.6 0.2     3.1 12.5 0.8 1.6 1.6 0.8 >200 0.4 0.2 3.1 6.3 0.8 3.1 IV 3.1 0.8 12.5 50     1.6 6.3 1.6 1.6 >200 3.1 0.8 6.3 100 0.8 3.1 V 0.8, 0.4, 3.1, 25, 1.6,     6.3, 1.6, 0.4, >200, 0.8, 1.6, 12.5 100, 1.6, >200,  0.8 0.4 12.5 50 1.6     6.3 0.8 0.4 >200 0.8 1.6 12.5 200 1.6 >200 VI 1.6 0.8 3.1 50 3.1 12.5     1.6 0.8 >200 0.8 3.1 25 >2006.3>200 VII 0.8, 0.8, 50, 50, 0.8, 3.1, 0.4,     0.4, >200, 0.4, 0.8, 6.3 200, 3.1, 100,  0.8 0.4 15 50 0.4 1.6 0.2 0.2     >200 0.2 0.8 3.1 100 0.8 100 VIII 0.4 0.1 1.6 6.3 0.8 1.6 1.6 0.4 >200     0.8 0.4 3.1 200 0.8 >200 IX 0.4 0.2 6.3 12.5 0.8 1.6 0.8 0.4 >200 0.4     0.2 1.6 100 0.8 200

                  TABLE 2                                                         ______________________________________                                               ED.sub.50 in vivo (mg./kg.)                                                   E. coli 12140                                                                              Kleb. pneumo. 4200                                        Compound s.c.     p.o.      s.c.     p.o.                                     ______________________________________                                        I        1.56     --        0.28     --                                       II       0.58      35       0.39     18.6                                     III      0.70      25       0.68     21.5                                     IV       18       >50       15.7     --                                       V        4        >50       0.26     --                                       VI       4.4      --        1.56     --                                       VII      0.86     --        0.24     --                                       VIII     1.32     >50       2        --                                       IX       1.56      44       --       --                                       ______________________________________                                    

                  TABLE 3                                                         ______________________________________                                        Compound Number                                                                           Compound Name                                                     ______________________________________                                        I           7-D-Mandelamido-3-(1-sulfomethyl-                                             tetrazol-5-ylthiomethyl)-3-cephem-                                            4-carboxylic acid                                                 II          7-D-Mandelamido-3-[1-(2-sulfoethyl)-                                          tetrazol-5-ylthiomethyl]-3-cephem-                                            4-carboxylic acid                                                 III         7-D-Mandelamido-3-[1-(2-sulfamoyl-                                            ethyl)tetrazol-5-ylthiomethyl]-                                               3-cephem-4-carboxylic acid                                        IV          7-D-Mandelamido-3-[1-(5-sulfopentyl)-                                         tetrazol-5-ylthiomethyl]-3-cephem-                                            4-carboxylic acid                                                 V           7-(2-Thienylacetamido)-3-(1-sulfo-                                            methyltetrazol-5-ylthiomethyl)-3-                                             cephem-4-carboxylic acid                                          VI          7-(2-Thienylacetamido)-3-[1-(2-                                               sulfoethyl)tetrazol-5-ylthio-                                                 methyl]-3-cephem-4-carboxylic acid                                VII         7-Trifluoromethylthioacetamido-                                               3-(1-sulfomethyltetrazol-5-ylthio-                                            methyl)-3-cephem-4-carboxylic acid                                VIII        7-(2-Thienylacetamido)-3-[1-(2-sulfamoyl-                                     ethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-                                    carboxylic acid                                                   IX          7-Trifluoromethylthioacetamido-3-                                             [1-(2-sulfamoylethyl)tetrazol-5-ylthio-                                       methyl]-3-cephem-4-carboxylic acid                                ______________________________________                                    

In addition, the active compounds of this invention exhibit broadspectrum activity and show advantageously high blood serum levels andhalf-life values.

Pharmaceutical compositions having antibacterial activity which comprisea pharmaceutical carrier containing an active but non-toxic quantity ofa compound of Formula I as well as methods of combatting bacterialinfections by administering such a composition to an infected host in anontoxic amount sufficient to combat such infections are also objects ofthis invention. The administration may be by parenteral injection suchas subcutaneously, intramuscularly or intravenously. The injection ofsuitably prepared sterile solutions or suspensions containing aneffective, nontoxic amount of the new cephalosporin compound is thepreferred route of administration.

The compounds of Formula I are formulated and administered in the samemanner as other cephalosporins. The dosage regimen comprisesadministration, preferably by injection, of an active but nontoxicquantity of a compound of Formula I selected from the dosage unit rangeof from 100 to 1000 mg. with the total daily dosage regimen being from400 mg. to 6 g. The precise dosages are dependent upon the age andweight of the subject and on the infection being treated and can bedetermined by those skilled in the art based on the data disclosedherein compared with that available to the art attained with knowncephalosporins.

Also considered within the scope of this invention are the 7α-methoxyanalogs of the compounds of Formula I, which compounds are representedby the following structural formula: ##STR7## or a non-toxicpharmaceutically acceptable salt thereof, in which R¹, R², R³ and n areas previously defined hereabove.

A selected group of the compounds of Formula IV are those where R¹ ishydrogen and n is one to five.

Representative of the compounds of Formula IV are7α-methoxy-7β-(2-thienylacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid,7α-methoxy-7β-trifluoromethylthioacetamido-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid,7β-D-mandelamido-7α-methoxy-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid,7α-methoxy-7β-(D-α-aminophenylacetamido)-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid and7β-D-mandelamido-7α-methoxy-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid.

When R² is hydroxy the compounds of Formula IV are preferably preparedby displacing the 3-acetoxy group from a7α-methoxy-7β-acylaminocephalosporanic acid or salt thereof, suitablyprotected as necessary, with a substituted tetrazole thiol of Formula IIwhere R² is hydroxy, or a corresponding salt, with subsequent removal ofthe protective group(s) and conversion of any salts to the correspondingfree acids, all as described hereinabove. The7α-methoxy-7β-acylaminocephalosporanic acids, salts or esters are eitherknown to the art or are prepared by known methods.

When R² is amino, lower alkylamino or di(lower)alkylamino, the compoundsof Formula IV are preferably prepared by acylation of a7β-amino-7α-methoxy-3-substituted tetrazolylthiomethylcephalosporinnucleus of Formula V: ##STR8## in which R¹, R⁴ and n are as previouslydefined hereabove and R² ' is amino, lower alkylamino ordi(lower)alkylamino with an appropriate acylating agent, suitablyprotected as necessary, by the procedures described above followed byremoval of the protective groups, when present, also as described above.The compounds of Formula V are also considered as objects of thisinvention.

The 7β-amino-7α-methoxy cephalosporin nuclei of Formula V are preparedby reaction of a 7-amino cephalosporin of Formula III wherein R² isamino, lower alkylamino or di(lower)alkylamino and R⁴ is a protectingester group such as a t-butyl group with3,5-di-t-butyl-4-hydroxybenzaldehyde with azeotropic removal of water.Subsequent treatment of the product thus formed with lead dioxide andreaction of the oxidized intermediate with methanol followed by cleavageof the imine function with, for example, Girard reagent T(trimethylaminoacetohydrazide chloride) followed by removal of theprotective group(s) as desired gives the corresponding compounds ofFormula V.

As with the compounds of Formula I, all non-toxic pharmaceuticallyacceptable salts and all isomers, including separated isomers andmixtures thereof, of the compounds represented by Formula IV areincluded within the scope of this invention.

The compounds of Formula IV have anti-bacterial activity against bothGram-positive and Gram-negative organisms. They are administered andformulated in the same manner as previously described for the compoundsof Formula I.

The following examples illustrate the invention, but are not to beconstrued as limiting the scope thereof. Temperatures are in degreesCentigrade unless otherwise stated.

EXAMPLE 17-D-Mandelamido-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

To a solution of 112 g. (2.0 mol.) of potassium hydroxide and 111 g.(1.0 mol.) of aminomethanesulfonic acid in 250 ml. of water at 25° wasadded 71 ml. of carbon disulfide. The reaction mixture was stirred for12 hours and 250 ml. of ethanol was added. The reaction vessel wasfitted with a reflux condenser and 62 ml. (1.0 mol.) of methyl iodidewas added. When the exothermic reaction cooled to ambient temperaturethe solid product was collected by filtration. The solid was extractedwith hot methanol and the extract was concentrated to give methylsulfomethyldithiocarbamate as the potassium salt.

A mixture of 45.3 g. (0.19 mol.) of methyl sulfomethyldithiocarbamatepotassium salt and 16.9 g. (0.26 mol.) of sodium azide in 425 ml. ofwater was heated at 80° for 4.75 hours. The reaction mixture was passedthrough an Amberlite IR-120H ion exchange resin column and eluted withwater until the pH of the eluant became 3.5. The eluant was extractedwith ether and the aqueous solution was evaporated to dryness to give1-sulfomethyltetrazole-5-thiol.

1-Sulfomethyltetrazole-5-thiol was dissolved in acetone and a 30%solution of sodium 2-ethylhexanoate in isopropanol was added.1-Sulfomethyltetrazole-5-thiol sodium salt precipitated and wascollected by filtration.

A mixture of 27.4 g. (0.062 mol.) of 7-D-mandelamidocephalosporanic acidmethanolate, 10.2 g. (0.047 mol.) of 1-sulfomethyltetrazole-5-thiolsodium salt and 9.2 g. (0.109 mol.) of sodium bicarbonate in 300 ml. ofwater was heated at 70° for one hour. The reaction mixture was cooled(ice bath) and acidified to pH 1.8 with 3N hydrochloric acid. Themixture was extracted with ethyl acetate, filtered and chromatographedon an Amberlite XAD-8 resin column with water containing increasingamounts of methanol as the eluant to give the title compound.

The title compound was dissolved in methanol and a 5% solution of sodiummethoxide in methanol was added until pH 7.0. Addition of ethanolprecipitated the product salt which was collected, dissolved in waterand lyophilized to give7-D-mandelamido-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid disodium salt.

    C.sub.18 H.sub.16 N.sub.6 O.sub.8 S.sub.3 . 2 Na . 1.75 H.sub.2 O

calculated: 34.98% C; 3.18% H; 13.59% N Found: 35.09% C; 3.17% H; 13.27%N

EXAMPLE 27-(D-α-Aminophenylacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

A solution of 7.58 g. (0.015 mol.) of7-(D-α-t-butoxycarbonylaminophenylacetamido)cephalosporanic acid, 1.96g. (0.01 mol.) of 1-sulfomethyltetrazole-5-thiol and 2.52 g. (0.03 mol.)of sodium bicarbonate in 125 ml. of water is stirred at 60° for fivehours while maintaining the pH at 7.0-7.2 by addition of sodiumbicarbonate. The mixture is cooled and extracted with ethyl acetate. Theaqueous phase is acidified to pH 2.5 with 3N hydrochloric acid and theacidic solution is extracted again with ethyl acetate. The aqueous phaseis brought to pH 7.1 by addition of 5% sodium carbonate solution, thenpassed through a XAD-4 ion exchange resin column and eluted with waterand methanol to give7-(D-α-t-butoxycarbonylaminophenylacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid disodium salt.

7-(D-α-t-butoxycarbonylaminophenylacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid disodium salt is stirred at 25° with 25 ml. of trifluoroacetic acidand 25 ml. of 1,3-dimethoxybenzene for 2.25 hours. The mixture isevaporated to dryness, ether is added to the residue and the precipitateis collected, washed with ether, stirred in acetonitrile for two hours,then collected and dried in vacuo to give the title compound.

EXAMPLE 37-D-Mandelamido-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

A solution of 2.73 g. (0.01 mol.) of 2-phthalimidoethanesulfonylchloride in 20 ml. of chloroform was added dropwise to a solution of2.19 g. (0.03 mol.) of t-butylamine in 20 ml. of chloroform at 5°. Thereaction mixture was warmed to ambient temperature and stirred for threehours. The precipitate was removed by filtration and the filtrate wasevaporated to dryness to give a residue which was purified bychromatography on silica with 19:1 chloroform-methanol as eluant to give2-N-t-butylphthalimidoethanesulfonamide.

2-N-t-Butylphthalimidoethanesulfonamide (2.10 g., 6.78 mmol.) wassuspended in 20 ml. of ethanol and 0.344 g. of hydrazine hydrate wasadded. The reaction mixture was refluxed for three hours, thenevaporated to dryness. The residue was suspended in 45 ml. of water andacidified to pH 3.0 by addition of dilute hydrochloric acid. The acidsolution was filtered and the filtrate evaporated to dryness to give2-aminoethane-N-t-butylsulfonamide hydrochloride.

2-Aminoethane-N-t-butylsulfonamide hydrochloride (1.25 g., 5.78 mmol.)was added to a solution of 1.17 g. (11.56 mmol.) of triethylamine in 20ml. of ethanol. Carbon disulfide (0.44 g., 5.78 mmol.) was added, themixture was stirred at 25° for 1.5 hours, then 0.82 g. (5.78 mmol.) ofmethyl iodide in 5 ml. of ethanol was added and the resulting mixturewas stirred for 1.5 hours. The mixture was evaporated to dryness and theresidue was dissolved in water and acidified to pH 2.0 with dilutehydrochloric acid. The aqueous mixture was extracted with ethyl acetateand the extract was dried (MgSO₄) and evaporated to dryness to givemethyl 2-(N-t-butylsulfamoyl)ethyldithiocarbamate.

Methyl 2-(N-t-butylsulfamoyl)ethyldithiocarbamate was treated withsodium azide as described in the procedure of Example 1 for 35 minutesto give 1-(2-N-t-butylsulfamoylethyl)tetrazole-5-thiol.

1-(2-N-t-Butylsulfamoylethyl)tetrazole-5-thiol (1.0 g.) was suspended in10 ml. of anisole and 20 ml. of trifluoroacetic acid is added. Thesolution was heated at 56° for 3.5 hours, then cooled. The precipitatewas collected by filtration and washed with petroleum ether to give1-(2-sulfamoylethyl)tetrazole-5-thiol.

A solution of 0.210 g. (2.5 mmol.) of sodium bicarbonate in 5 ml. ofwater was added to a suspension of 0.272 g. (1 mmol.) of7-aminocephalosporanic acid in 5 ml. of water and 2.5 ml. of acetone at15°. The solution was heated to 45°, a solution of 0.314 g. (1.5 mmol.)of 1-(2-sulfamoylethyl)tetrazole-5-thiol in 10 ml. of acetone was addedand the reaction mixture was refluxed for two hours while maintainingthe pH at 7.4-7.6 by addition of aqueous sodium bicarbonate solution.The mixture was cooled and acidified to pH 4.0 with dilute hydrochloricacid. The precipitate was collected by filtration to give7-amino-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid.

To a solution of 1.26 g. (15 mmol.) of sodium bicarbonate in 75 ml. ofacetone and 50 ml. of water at 5° was added 2.1 g. (5 mmol.) of7-amino-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid. The solution was cooled to -10° and a solution of 1.55 g. (5.5mmol.) of D-O-dichloroacetylmandeloyl chloride in 25 ml. of acetone wasadded. The reaction mixture was stirred for 30 minutes in the cold whilemaintaining the pH at 7.2 by addition of aqueous sodium bicarbonate,then for 1.5 hours at 25°. The mixture was extracted with ether and theaqueous phase was brought to pH 9.3 with 5% sodium carbonate and stirredfor 1.5 hours at 25°. The aqueous mixture was extracted with ether, thepH adjusted to 4.5 and the solution was again extracted with ether. Theaqueous phase was acidified to pH 1.5 with dilute hydrochloric acid andit was extracted with ethyl acetate. Evaporation of the extract todryness gave a solid which was suspended in ethyl acetate and filtered.The filtrate was diluted with ether and petroleum ether to precipitatethe title compound.

EXAMPLE 47-D-Mandelamido-3-[1-(2-N-t-butylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

When an equivalent amount of1-(2-N-t-butylsulfamoylethyl)tetrazole-5-thiol is reacted with7-aminocephalosporanic acid as described in Example 3,7-amino-3-[1-(2-N-t-butylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid is obtained.

Acylation of7-amino-3-[1-(2-N-t-butylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid with D-O-dichloroacetylmandeloyl chloride according to theprocedure of Example 3 gives the title compound.

EXAMPLE 57-D-Mandelamido-3-[1-(2-N-methylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

Use of methylamine in the reaction with 2-phthalimidoethanesulfonylchloride described in Example 3,

followed by the subsequent synthetic steps described therein gives1-(2-N-methylsulfamoylethyl)tetrazole-5-thiol.

When 1-(2-N-methylsulfamoylethyl)tetrazole-5-thiol is reacted with7-aminocephalosporanic acid as described in Example 3 and the product7-amino-3-[1-(2-N-methylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid is acylated with D-O dichloroacetylmandeloyl chloride as describedtherein, the title compound is obtained.

EXAMPLE 67-(D-α-Amino-4-hydroxyphenylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

A solution of 7.82 g. (0.015 mol.) of7-(D-α-t-butoxycarbonyl-4-hydroxyphenylacetamido)cephalosporanic acid,4.6 g. (0.022 mol.) of 1-(2-sulfamoylethyl)tetrazole-5-thiol and sodiumbicarbonate are reacted according to the procedure described in Example2. After cooling, the reaction mixture is extracted with ethyl acetate.Fresh ethyl acetate is added to the aqueous phase and it is acidifiedwith stirring to pH 2.8 with 6N sulfuric acid. The layers are separatedand the aqueous phase is again extracted with ethyl acetate. Thecombined extracts are washed with water, dried (MgSO₄) and the solventevaporated to give7-D-α-t-butoxycarbonyl-4-hydroxyphenylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid.

7-(D-α-t-Butoxycarbonyl-4-hydroxyphenylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid is treated with trifluoroacetic acid as described in Example 2 togive the title compound.

EXAMPLE 77-D-Mandelamido-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

2-Aminoethanesulfonic acid (50 g., 0.4 mol.) was added to a solution of45 g. (0.8 mol.) of potassium hydroxide in 100 ml. of water at 25°.Carbon disulfide (24.4 ml., 0.4 mol.) was added and the reaction mixturewas refluxed for 2.5 hours. Ethanol was added to the warm solution, themixture was cooled to ambient temperature, 57 g. (0.4 mol.) of methyliodide was added and the resulting mixture was stirred for 1.5 hours.The mixture was evaporated in vacuo and the residue recrystallized fromhot ethanol containing 3% water to give methyl2-sulfoethyldithiocarbamate potassium salt.

A mixture of 21.5 g. (0.087 mol.) of methyl 2-sulfoethyldithiocarbamatepotassium salt (0.5 hydrate) and 7.16 g. (0.11 mol.) of sodium azide in200 ml. of water was refluxed for two hours. The solution was cooled to25° and extracted with ethyl acetate. The aqueous phase was treated withAmberlite IR-120H resin, washed with ether and evaporated to give anoil. The oil was dissolved in acetone, the solution was filtered and thefiltrate was evaporated to dryness to give1-(2-sulfoethyl)-tetrazole-5-thiol. The thiol was dissolved inisopropanol, cyclohexylamine was added until pH 8-9 and acetonitrile wasadded to give 1-(2-sulfoethyl)tetrazole-5-thiol as thedi-cyclohexylamine salt.

1-(2-Sulfoethyl)tetrazole-5-thiol di-cyclohexylamine salt was dissolvedin water and treated with Amberlite IR-120H resin to give1-(2-sulfoethyl)tetrazole-5-thiol.

To a solution of 2.1 g. (0.01 mol.) of 1-(2-sulfoethyl)tetrazole-5-thiol in 100 ml. of water was added 6.4 g. (0.015 mol.) of7-D-mandelamidocephalosporanic acid sodium salt and 1.68 g. (0.02 mol.)of sodium bicarbonate. The mixture was stirred at 70° for 2.5 hours thencooled and acidified to pH 1.8 with 3N hydrochloric acid. The acidsolution was extracted with ethyl acetate and ether, acidified to pH 0.9and chromatographed on a XAD-8 resin column with water as eluant to givethe title compound.

The title compound was converted to the corresponding disodium salt bytreatment with sodium methoxide as described in the procedure of Example1.

    C.sub.19 H.sub.18 N.sub.6 O.sub.8 S.sub.3 . 2 Na . 2 H.sub.2 O

calculated: 35.85% C; 3.48% H; 13.20% N Found: 36.21% C; 3.29% H; 12.96%N

EXAMPLE 87-D-Mandelamido-3-(1-sulfamoylmethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

A suspension of 15.1 g. (0.136 mol.) of aminomethanesulfonic acid and14.2 g. (0.145 mol.) of anhydrous potassium acetate in 48 ml. of aceticacid is refluxed for ten minutes. Phthalic anhydride (21.4 g., 0.145mol.) is then added and the resulting mixture is refluxed for 2.5 hours.The product is collected by filtration and washed with acetic acid andethanol to give phthalimidomethanesulfonic acid potassium salt.

To 41.7 g. (0.15 mol.) of phthalimidomethanesulfonic acid potassium saltin 220 ml. of dry benzene is added 22.5 g. (0.132 mol.) of phosphoruspentachloride. The reaction mixture is refluxed on a steam bath for onehour, then an additional 22.5 g. of phosphorus pentachloride is addedand heating is continued for 1.5 hours. The reaction mixture isevaporated to dryness, crushed ice is added to the residue and theslurry is filtered. The product is washed with water to givephthalimidomethanesulfonyl chloride.

When phthalimidomethanesulfonyl chloride is substituted in the procedureof Example 3 for 2-phthalimidoethanesulfonyl chloride,N-t-butylphthalimidomethanesulfonamide is prepared which is converted to1-N-t-butylsulfamoylmethyltetrazole-5-thiol as described therein.Treatment of 1-N-t-butylsulfamoylmethyltetrazole-5-thiol withtrifluoroacetic acid as described in Example 3 gives1-sulfamoylmethyltetrazole-5-thiol.

Reaction of 1-sulfamoylmerthyltetrazole-5-thiol with7-aminocephalosporanic acid and treatment of the product7-amino-3-(1-sulfamoylmethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid with D-O-dichloroacetylmandeloyl chloride as described in Example 3gives the title compound.

EXAMPLE 97-(D-α-Amino-4-hydroxyphenylacetamido)-3-[1-(2-N,N-dimethylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

1-(2-N,N-Dimethylsulfamoylethyl)tetrazole-5-thiol is prepared fromreaction of dimethylamine and 2-phthalimidoethanesulfonyl chloridefollowed by conversion of the product thus obtained to the tetrazolethiol by the reaction sequence described in the procedure of Example 3.

When 1-(2-N,N-dimethylsulfamoylethyl)tetrazole-5-thiol is reacted with7-(D-α-t-butoxycarbonyl-4-hydroxyphenylacetamido)cephalosporanic acid asdescribed in Example 6 and the product is de-blocked as described above,the title compound is obtained.

EXAMPLE 107-D-Mandelamido-3-[1-(3-sulfopropyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

When an equivalent amount of 3-aminopropanesulfonic acid was substitutedin the procedure of Example 7 for 2-aminoethanesulfonic acid, methyl3-sulfopropyldithiocarbamate potassium salt was prepared.

Reaction of methyl 3-sulfopropyldithiocarbamate potassium salt withsodium azide as described in Example 7 gave1-(3-sulfopropyl)tetrazole-5-thiol.

Substitution of an equivalent amount of1-(3-sulfopropyl)tetrazole-5-thiol in the procedure of Example 7 inplace of 1-(2-sulfoethyl)tetrazole-5-thiol in the reaction with7-D-mandelamidocephalosporanic acid sodium salt gives the titlecompound.

The title compound is converted to the corresponding sodium salt asdescribed in the procedure of Example 1.

EXAMPLE 117-D-Mandelamido-3-[1-(3-sulfamoylpropyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

When 3-phthalimidopropanesulfonyl chloride is substituted in theprocedure of Example 3 for 2-phthalimidoethanesulfonyl chloride,3-N-t-butylphthalimidopropanesulfonamide is prepared, which is convertedto 1-(3-N-t-butylsulfamoylpropyl)tetrazole-5-thiol as described therein.Treatment of 1-(3-N-t-butylsulfamoylpropyl)-tetrazole-5-thiol withtrifluoroacetic acid as described in Example 3 gives1-(3-sulfamoylpropyl)tetrazole-5-thiol.

Reaction of 1-(3-sulfamoylpropyl)tetrazole-5-thiolwith7-aminocephalosporanic acid and treatment of the resulting7-amino-3-[1-(3-sulfamoylpropyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid with D-O-dichloroacetylmandeloyl chloride as described in Example 3gives the title compound.

EXAMPLE 127-D-Mandelamido-3-[1-(5-sulfopentyl)tetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

Substitution of an equivalent amount of 5-aminopentanesulfonic acid inthe procedure of Example 7 for 2-aminoethanesulfonic acid gave methyl5-sulfopentyldithiocarbamate potassium salt.

Reaction of methyl 5-sulfopentyldithiocarbamate potassium salt withsodium azide as described in Example 7 gave1-(5-sulfopentyl)tetrazole-5-thiol.

A mixture of 4.55 g. (0.010 mol.) of 7-D-mandelamidocephalosporanic acidsodium salt and 3.25 g. (0.011 mol.) of1-(5-sulfopentyl)tetrazole-5-thiol disodium salt, prepated as previouslydescribed, in 80 ml. of water at pH 7.2 (adjusted by addition of sodiumbicarbonate) was heated at 65° for 4.5 hours. The reaction mixture wascooled, acidified to pH 1.6 with 3N hydrochloric acid and extracted withethyl acetate. The pH of the aqueous phase was brought to 7.0 byaddition of sodium bicarbonate and the solution was chromatographed on aXAD-4 resin column eluting with water and then methanol. The product wasdissolved in methanol and ethanol was added to the solution toprecipitate the title compound as its disodium salt.

    C.sub.22 H.sub.24 N.sub.6 O.sub.8 S.sub.3 . 2 Na . 1.5 H.sub.2 O . 0.75 C.sub.2 H.sub.6 O

calculated: 40.08% C; 4.50% H; 11.93% N; 13.65% S Found: 40.01% C; 4.20%H; 10.71% N; 13.54% S

7-D-Mandelamido-3-[1-(5-sulfopentyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid disodium salt is converted to the title compound as describedherein.

EXAMPLE 137-D-Mandelamido-3-[1-(5-sulfamoylpentyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

Use of 5-aminopentanesulfonic acid in the procedure of Example 8 inplace of aminomethanesulfonic acid followed by reaction of the productthus obtained with phosphorus pentachloride gives5-phthalimidopentanesulfonyl chloride.

When 5-phthalimidopentanesulfonyl chloride is used as a startingmaterial in the sequence described in Example 3,1-(5-t-butylsulfamoylpentyl)tetrazole-5-thiol is obtained. Treatment of1-(5-N-t-butylsulfamoylpentyl)-tetrazole-5-thiol with trifluoroaceticacid as described above gives 1-(5-sulfamoylpentyl)tetrazole-5-thiol.

Reaction of 1-(5-sulfamoylpentyl)tetrazole-5-thiol with7-aminocephalosporanic acid and treatment of the resulting7-amino-3-[1-(5-sulfamoylpentyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid with D-O-dichloroacetylmandeloyl chloride as described in Example 3gives the title compound.

EXAMPLE 147-Trifluoromethylthioacetamido-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

A solution of 2.18 g. (10.0 mmol.) of 1-sulfomethyltetrazole-5-thiolsodium salt, prepared as described above, 0.840 g. of sodium bicarbonateand 5.45 g. (12.5 mmol.) of7-trifluoromethylthioacetamidocephalosporanic acid sodium salt in 60 ml.of water was stirred at 70-75° for five hours while maintaining the pHat 6.8 by addition of 5% aqueous sodium carbonate solution. The reactionmixture was cooled and diluted with water. Ethyl acetage was added andthe mixture was acidified to pH 2.0 with 6N hydrochloric acid. Theaqueous phase was extracted with ethyl acetate then brought to pH 6.8 byaddition of sodium bicarbonate and chromatographed on XAD-4 resin withwater and methanol as eluants. The product-containing fractions wereevaporated to dryness to give a residue which was dissolved in water andlyophilized to give the title compound as the disodium salt.

    C.sub.13 H.sub.11 F.sub.3 N.sub.6 O.sub.7 S.sub.4 . 2 Na . 2.25 H.sub.2 O

calculated: 24.59% C; 2.46% H; 13.23% N Found: 24.54% C; 2.18% H; 12.85%N

An aqueous solution of 7-trifluoromethylthioacetamido-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid disodium salt is treated with Amberlite IR-120H ion exchange resinas described above to give the title compound.

EXAMPLE 157-Trifluoromethylthioacetamido-3-[1-(3-sulfamoylpropyl)tetrazol-5-ylthiomethyl]-3-cephem-4carboxylicacid

7-Trifluoromethylthioacetamido-3-[1-(3-sulfamoylpropyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid sodium salt is prepared by substitution of an equivalent amount of1-(3-sulfamoylpropyl)tetrazole-5-thiol sodium salt, prepared asdescribed above, in the procedure of Example 14.

The title compound is obtained from the salt as described above.

EXAMPLE 167-Trifluoromethylthioacetamido-3-[1-(2-sulfamoylethyl)-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

A mixture of 2.18 g. (5 mmol.) of7-trifluoromethylthioacetamidocephalosporanic acid sodium salt and 1.75g. (8 mmol.) of 1-(2-sulfamoylethyl)tetrazole-5-thiol in 50 ml. of watermaintained at pH 7.0 by addition of sodium bicarbonate was heated at60-70° for three hours. The reaction mixture was cooled, acidified to pH3.5 and extracted with ethyl acetate. The extract was washed with water,dried (MgSO₄) and evaporated to dryness to give a residue which wasdissolved in ethyl acetate. Ether was added, the solution was filtered,petroleum ether was added to the filtrate and the resulting precipitateswas collected by filtration and dissolved in methanol. A 5% solution ofsodium methoxide in methanol was added to the methanol solution until pH7.1. Ether was added and the precipitate was collected and dried invacuo to give7-trifluoromethylthioacetamido-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid sodium salt.

7-Trifluoromethylthioacetamido-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid sodium salt is converted to the title compound by methods describedabove.

EXAMPLE 177-D-Mandelamido-3-[1-(10-sulfodecyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

A suspension of 56 g. (1.0 mol.) of potassium hydroxide and 118.7 g.(0.5 mol.) of 10-aminodecanesulfonic acid in 170 ml. of water is stirredfor 30 minutes at 25°, then 40 g. (0.52 mol.) of carbon disulfide and 80ml. of ethanol are added and the reaction mixture is stirred at 25° for12 hours. The mixture is refluxed gently for two hours and cooled.Methyl iodide (71 g., 0.3 mol.) and 130 ml. of ethanol are added to themixture and it is stirred at 25° for 12 hours. The mixture is evaporatedto remove the ethanol and the solid residue is collected by filtrationto give methyl 10-sulfodecyldithiocarbamate.

Methyl 10-sulfodecyldithiocarbamate (31.4 g., 0.096 mol.) is reactedwith 6.5 g. (0.1 mol.) of sodium azide as described above to give1-(10-sulfodecyl)-tetrzole-5-thiol.

1-(10-Sulfodecyl)tetrazole-5-thiol (4.84 g., 15 mmol.) is slowly addedto a solution of 3.36 g. (40 mmol.) of sodium bicarbonate in 100 ml. ofwater. 7-D-Mandelamidocephalosporanic acid (4.20 g., 10 mmol.) is thenadded and the mixture is heated at 65° for 3.5 hours. The mixture isfiltered, the filtrate is extracted with ethyl acetate and the aqueouslayer is acidified to pH 4 and extracted again with ethyl acetate. Theextract is dried (MgSO₄) and evaporated to dryness to give the titlecompound.

EXAMPLE 187-D-Mandelamido-3-[1-(10-sulfamoyldecyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

Use of 10-aminodecanesulfonic acid in the procedure of Example 8 inplace of aminomethanesulfonic acid followed by reaction of the productthus obtained with phosphorus pentachloride gives10-phthalimidodecanesulfonyl chloride.

When 10-phthalimidodecanesulfonyl chloride is used as a startingmaterial in the sequence described in Example 3,1-(10-N-t-butylsulfamoyldecyl)tetrazole-5-thiol is obtained. Treatmentof 1-(10-N-t-butylsulfamoyldecyl)-tetrazole-5-thiol with trifluoroaceticacid gives 1-(10-sulfamoyldecyl)tetrazole-5-thiol.

Reaction of 1-(10-sulfamoyldecyl)tetrazole-5-thiol with7-aminocephalosporanic acid and treatment of the resulting7-amino-3-[1-(10-sulfamoyldecyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid with D-O-dichloroacetylmandeloyl chloride as described in Example 3gives the title compound.

EXAMPLE 197-D-Mandelmido-3-[1-(2-sulfo-1-methylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

Substitution of an equivalent amount of 2-aminopropanesulfonic acid inthe procedure of Example 7 for 2-aminoethanesulfonic acid gives methyl(2-sulfo-1-methyl)-ethyldithiocarbamate potassium salt.

Treatment of methyl (2-sulfo-1-methyl)ethyldithiocarbamate potassiumsalt with sodium azide also as described in Example 7 gives1-(2-sulfo-1-methylethyl)-tetrazole-5-thiol.

7-D-Mandelamidocephalosporanic acid and1-(2-sulfo-1-methylethyl)tetrazole-5-thiol are reacted in the presenceof excess sodium bicarbonate as described in Example 7 to give the titlecompound.

EXAMPLE 20

When an equivalent amount of aminosulfonic acid listed below:

2-aminobutanesulfonic acid

1-amino-2-methylpropanesulfonic acid

1-amino-3-methylbutanesulfonic acid

is used in the procedure of Example 7 in place of 2-aminomethanesulfonicacid and the resulting dithiocarbamates are treated with sodium azide asdescribed therein, the following substituted tetrazole thiols areobtained:

1-(1-sulfomethylpropyl)tetrazole-5-thiol

1-(2-methyl-1-sulfopropyl)tetrazole-5-thiol

1-(3-methyl-1-sulfobutyl)tetrazole-5-thiol.

Reaction of a tetrazole thiol listed above with7-D-mandelamidocephalosporanic acid as described hereinabove gives thefollowing compounds of this invention:

7-D-mandelamido-3-[1-(1sulfomethylpropyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-D-mandelamido-3-[1-(2-methyl-1-sulfopropyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-D-mandelamido-3-[1-(3-methyl-1-sulfobutyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid.

Likewise, reaction of a substituted tetrazole thiol listed above with7-(D-α-t-butoxycarbonyl-4-hydroxyphenylacetamido)cephalosporanic acid or7-trifluoromethylthioacetamidocephalosporanic acid according to theprocedures described herein within subsequent removal of the protectivegroups as necessary, gives the corresponding7-(D-α-amino-4-hydroxyphenylacetamido)-3-(sulfoalkyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacids and7-trifluoromethylthioacetamido-3-(sulfoalkyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacids.

EXAMPLE 21

When an aminosulfonic acid listed below:

2-aminopropanesulfonic acid

2-aminobutanesulfonic acid

1-amino-2-methylpropanesulfonic acid

1-amino-3-methylbutanesulfonic acid is substituted as a startingmaterial in the procedure of Example 8 for aminomethanesulfonic acid,the subsequent steps described therein and in Example 3 are carried outand the resulting N-t-butyl sulfamoyl tetrazoles are treated withtrifluoroacetic acid as described therein, the following substitutedtetrazole thiols are obtained:

1-(1-methyl-2-sulfamoylethyl)tetrazole-5-thiol

1-(1-sulfamoylmethylpropyl)tetrazole-5-thiol

1-(2-methyl-1-sulfamoylpropyl)tetrazole-5-thiol

1-(3-methyl-1-sulfamoylbutyl)tetrazole-5-thiol.

Reaction of a tetrazole thiol listed above with 7-aminocephalosporanicacid followed by treatment of the resulting 7-amino-3-(sulfamoylalkyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid withD-O-dichloroacetylmandeloyl chloride as described in Example 3 gives thecompounds of this invention listed below:

7-D-mandelamido-3-[1-(1-methyl-2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-D-mandelamido-3-[1-(1-sulfamoylmethylpropyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-D-mandelamido-3-[1-(2-methyl-1-sulfamoylpropyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-D-mandelamido-3-[1-(3-methyl-1-sulfamoylbutyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid.

EXAMPLE 22

Reaction of the N-t-butoxycarbonyl derivative of the followingcephalosporanic acids:

7-(α-amino-4-formamidophenylacetamido)cephalosporanic acid

7-(α-amino-3-formamidophenylacetamido)cephalosporanic acid

7-(α-amino-4-ureidophenylacetamido)cephalosporanic acid

7-(α-amino-3-ureidophenylacetamido)cephalosporanic acid

7-(α-amino-4-hydroxymethylphenylacetamido)cephalosporanic acid

7-(α-amino-1,4-cyclohexadienylacetamido(cephalosporanic acid

7-(α-amino-3-fluoro-4-hydroxyphenylacetamido)cephalosporanic acid with1-(2-sulfamoylethyl)tetrazole-5-thiol as described in the procedure ofExample 2 followed by removal of the protective group as describedtherein gives the following compounds of this invention:

7-(α-amino-4-formamidophenylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(α-amino-3-formamidophenylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(α-amino-4-ureidophenylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(α-amino-3-ureidophenylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(α-amino-4-hydroxymethylphenylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(α-amino-1,4-cyclohexadienylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(α-amino-3-fluoro-4-hydroxyphenylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid.

By similar procedures described hereinabove, 7-(α-amino substitutedphenylacetamido)-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacids are prepared by reaction of the N-t-butoxycarbonyl derivative ofthe cephalosporanic acids listed above with1-(2-sulfoethyl)tetrazole-5-thiol followed by removal of the protectivegroup as previously described.

EXAMPLE 23

7-(4-Hydroxymandelamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid is prepared by reaction of 7-(4-hydroxymandelamido)cephalosporanicacid and 1-(2-sulfamoylethyl)tetrazole-5-thiol as described in theprocedure of Example 3.

Likewise, when 7-(4-hydroxymandelamido)cephalosporanic acid sodium saltis substituted in the procedure of Example 7 for7-D-mandelamidocephalosporanic acid sodium salt,7-(4-hydroxymandelamido)-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid is obtained.

EXAMPLE 24

Reaction of a cephalosporanic acid listed below:

7-(α-hydroxy-2-thienylacetamido)cephalosporanic acid

7-(α-carboxy-2-thienylacetamido)cephalosporanic acid

7-(α-sulfophenylacetamido)cephalosporanic acid with1-(2-sulfamoylethyl)tetrazole-5-thiol or1-(2-sulfoethyl)tetrazole-5-thiol as described hereinabove gives thefollowing compounds of this invention:

7-(α-hydroxy-2-thienylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(α-hydroxy-2-thienylacetamido)-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(α-carboxy-2-thienylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(α-carboxy-2-thienylacetamido)-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(α-sulfophenylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(α-sulfophenylacetamido)-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid.

EXAMPLE 25

When the N-t-butoxycarbonyl derivative of7-(α-amino-2-thienylacetamido)cephalosporanic acid is reacted with1-(2-sulfamoylethyl)tetrazole-5-thiol or1-(2-sulfoethyl)tetrazole-5-thiol followed by removal of the protectivegroups as described above,7-(α-amino-2-thienylacetamido)-3-[1-(2-sulfamoylethy)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid and7-(α-amino-2-thienylacetamido)-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid are obtained, respectively.

EXAMPLE 26

A mixture of 2.5 g. (7.7 mmol.) ofD-α-(N-t-butoxycarbonyl)-4-aminophenylglycine, 1.8 g. (9.2 mmol.) ofα-bromoacetic acid t-butyl ester and 2.5 g. (19.2 mmol.) ofN,N-disopropylethylamine in 15 ml. of ethanol was stirred at 25° for 48hours. The solvent was removed in vacuo, the residue was diluted withethyl acetate and sodium bicarbonate and the pH was adjusted to 2.5. Thelayers were separated and the aqueous phase was again extracted withethyl acetate. The combined extracts were washed with saturated sodiumchloride solution, dried (MgSO₄) and evaporated to dryness to giveD-α-(N-t-butoxycarbonyl)-4-t-butoxycarbonylmethylaminophenylglycine.

A solution of 0.380 g. (1.0 mmol.) ofD-α-(N-t-butoxycarbonyl)-4-t-butoxycarbonylmethylaminophenylglycine,0.296 g. (1.0 mmol.) of 7-aminocephalosporanic acid t-butyl ester and0.210 g. (1.0 mmole.) of dicyclohexylcarbodiimide in 25 ml. of 9:1 ethylacetate-methylene chloride was stirred at 0° for one hour. The reactionmixture was filtered and the filtrate was washed with 2.5% sulfuricacid, 5% sodium bicarbonate and water, dried (MgSO₄) and evaporated todryness to give7-(D-α-t-butoxycarbonylamino-4-carboxymethylaminophenylacetamido)cephalosporanicacid t-butyl ester. Deblocking was accomplished by stirring a mixture ofthe cephalosporanic acid t-butyl ester and 2 ml. of benzenethiol in 10ml. of trifluoroacetic acid at 25° for one hour. Evaporation of thereaction mixture to dryness gave7-(D-α-amino-4-carboxymethylaminophenylacetamido)cephalosporanic acid.

Reaction of the N-t-butoxycarbonyl derivative of7-(D-α-amino-4-carboxymethylaminophenylacetamido)cephalosporanic acidwith 1-(2-sulfamoylethyl)tetrazole-5-thiol or1-(2-sulfoethyl)tetrazole-5-thiol according to procedures describedhereinabove with subsequent removal of the protective group also asdescribed above gives7-(D-α-amino-4-carboxymethylaminophenylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl[-3-cephem-4-carboxylicacid and7-(D-α-amino-4-carboxymethylaminophenylacetamido)-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid, respectively.

EXAMPLE 27

When ethylamine or propylamine is used in the reaction with2-phthalimidoethanesulfonyl chloride described in the procedure ofExample 3 and the product formed is subjected to the synthetic stepsdescribed therein, the following tetrazole thiols are prepared:

1-(2-N-ethylsulfamoylethyl)tetrazole-5-thiol

1-(2-N-propylsulfamoylethyl)tetrazole-5-thiol.

Reaction of a tetrazole thiol listed above with 7-aminocephalosporanicacid, 7-D-mandelamidocephalosporanic acid,7-(D-α-t-butoxycarbonylamino-4-hydroxyphenylacetamido)cephalosporanicacid, 7-(2-thienylacetamido)cephalosporanic acid and7-trifluoromethylthioacetamidocephalosporanic acid, respectively, withremoval of the protective groups when necessary, as described above,gives the following compounds of this invention:

7-amino-3-[1-(2-N-ethylsulfamoylethyl)tetrazol-5-ylthiomethyl[-3-cephem-4-carboxylicacid

7-amino-3-[1-(2-N-propylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-D-mandelamido-3-[1-(2-N-ethylsulfamoylethyl)-tetrazol-5-ylthiomethyl[-3-cephem-4-carboxylic acid

7-D-mandelamido-3-]1-(2-N-propylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(D-α-amino-4-hydroxyphenylacetamido)-3-[1-(2-N-ethylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(D-α-amino-4-hydroxyphenylacetamido)-3-[1-(2-N-propylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(2-thienylacetamido)-3-[1-(2-N-ethylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(2-thienylacetamido)-3-[1-(2-N-propylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-trifluoromethylthioacetamido-3-[1-(2-N-ethylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-trifluoromethylthioacetamido-3-[1-(2-N-propylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid.

EXAMPLE 28

When diethylamine, dipropylamine or dibutylamine is substituted fordimethylamine in the procedure of Example 9, the following tetrazolthiols are prepared:

1-(2-N,N-diethylsulfamoylethyl)tetrazole-5-thiol

1-(2-N,N-dipropylsulfamoylethyl)tetrazole-5-thiol

1-(2-N,N-dibutylsulfamoylethyl)tetrazole-5-thiol.

Reaction of a tetrazole thiol listed above with 7-aminocephalosporanicacid, 7-D-mandelamidocephalosporanic acid,7-(D-α-t-butoxycarbonylamino-4-hydroxyphenylacetamido)-cephalosporanicacid, 7-(2-thienylacetamido)cephalosporanic acid and7-trifluoromethylthioacetamidocephalosporanic acid, respectively, withremoval of the protective groups when necessary, as described above,gives the following compounds of this invention:

7-amino-3-[1-(2-N,N-diethylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-amino-3-[1-(2-N,N-dipropylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-amino-3-[1-(2-N,N-dibutylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-D-mandelamido-3-[1-(2-N,N-diethylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-D-mandelamido-3-[1-(2-N,N-dipropylsulfamoylethyl)tetrazol-5-ythiomethyl]-3-cephem-4-carboxylicacid

7-D-mandelamido-3-[1-(2-N,N-dibutylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(D-α-amino-4-hydroxyphenylacetamido)-3-[1-(2-N,N-diethylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(D-α-amino-4-hydroxyphenylacetamido)-3-[1-(2-N,N-dipropylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(D-α-amino-4-hydroxyphenylacetamido)-3-[1-(2-N,N-dibutylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(2-thienylacetamido)-3-[1-(2-N,N-diethylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(2-thienylacetamido)-3-[1-(2-N,N-dipropylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(2-thienylacetamido)-3-[1-(2-N,N-dibutylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-trifluoromethylthioacetamido-3-[1-(2-N,N-diethylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-trifluoromethylthioacetamido-3-[1-(2-N,N-dipropylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-trifluoromethylthioacetamido-3-[1-(2-N,N-dibutylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid.

EXAMPLE 29

When a cephalosporanic acid listed below:

7-(1-tetrazolylacetamido)cephalosporanic acid

7-(3-sydnoneacetamido)cephalosporanic acid

7-(2-aminomethylphenylacetamido)cephalosporanic acid

is reacted with 1-(2-sulfamoylethyl)tetrazole-5-thiol by the proceduredescribed in Example 3, the following compounds of this invention areobtained, respectively:

7-(1-tetrazolylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(3-sydnoneacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(2-aminomethylphenylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid.

Similarly, reaction of the sodium salt of a cephalosporanic acid listedabove with 1-(2-sulfoethyl)tetrazole-5-thiol as described in Example 7gives the following compounds of this invention:

7-(1-tetrazolylacetamido)-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(3-sydnoneacetamido)-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-(2-aminomethylphenylacetamido)-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid.

EXAMPLE 30

Reaction of the sodium salt of a cephalosporanic acid listed below:

7-(2,2,2-trifluoroethylthioacetamido)-cephalosporanic acid

7-methylthioacetamidocephalosporanic acid

7-n-propylthioacetamidocephalosporanic acid

with 1-sulfomethyltetrazole-5-thiol sodium salt as described in theprocedure of Example 14 gives the following compounds of this inventionas final products:

7-(2,2,2-trifluoroethylthioacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

7-methylthioacetamido-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

7-n-propylacetamido-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid.

EXAMPLE 31

Reaction of the sodium salt of a cephalosporanic acid listed in Example30 with 1-(2-sulfamoylethyl)tetrazole-5-thiol sodium salt by theprocedure of Example 14 gives the 7-substituted-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacids listed below:

7-(2,2,2-trifluoroethylthioacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

    C.sub.15 H.sub.17 F.sub.3 N.sub.7 O.sub.6 S.sub.4  · Na ·0.5 CH.sub.4 O

calculated: 30.24% C; 3.11% H; 15.92% N Found: 30.62% C; 3.00% H; 15.32%N

7-methylthioacetamido-3[1-(2-sulfamoylethyl)-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

7-n-propylthioacetamido-3-[1-(2-sulfamoylethyl)tetrazol-5ylthiomethyl]-3-cephem-4-carboxylicacid.

EXAMPLE 327-(2,2,2-Trifluoroethylsulfinylacetamido)-3-(2-sulfamoylethyl)tetrazol-5ylthiomethyl-3-cephem-4-carboxylicacid

To a stirred solution of 5.7 g. (0.03 mol.) of2,2,2-trifluoroethylsulfinylacetic acid and 3.45 g. (0.03 mol.) ofN-hydroxysuccinimide in 50 ml. of tetrahydrofuran at 0° is added 6.2 g.(0.031 mol.) of dicyclohexylcarbodiimide. The reaction mixture isstirred at 0° for one hour than at 25° for 12 hours. The precipitate isfiltered and washed with tetrahydrofuran and the filtrate is evaporatedto dryness to give the activated ester of2,2,2-trifluoroethylsulfinylacetic acid.

A suspension of 4.17 g. (0.01 mol.) of7-amino-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid in 50 ml. of dry dimethylformamide is treated with 2 ml. oftriethylamine and the mixture is strirred for 15 minutes at 25° . Aslight excess of 0.01 mol. of the activated ester of2,2,2-trifluoroethylsulfinylacetic acid is added to the mixture and itis stirred an additional hour. The reaction mixture is evaporated todryness and water and ethyl acetate are added to the residue. The layersare separated, ethyl acetate is added to the aqueous phase and it isacidified to pH 2.5 by addition of 6N hydrochloric acid. The mixture isfiltered, the layers are separated and the aqueous phase is extractedwith ethyl acetate. The extract is washed with water, dried (MgSO₄) andevaporated to dryness to give the title compound.

EXAMPLE 337-(2,2,2-Trifluoroethylsulfonylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

To a solution of 9.0 g. (0.019 mol.) of7-amino-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid t-butyl ester and 3.9 g. (0.019 mol.) of2,2,2-trifluoroethylsulfonylacetic acid in tetrahydrofuran is addeddropwise a solution of 3.9 g. (0.019 mol.) of dicyclohexylcarbodiimidein 100 ml. of tetrahydrofuran. The reaction mixture is stirred at 25°for 12 hours, then filtered and concentrated to about 10 ml. The residueis filtered and evaporated to dryness to give7-(2,2,2-trifluoroethylsulfonylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid t-butyl ester.

The ester is dissolved in acetonitrile and trifluoroacetic acid isadded. The solution is stirred for three hours, then evaporated todryness to give the title compound.

EXAMPLE 347-Methylthioacetamido-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

To a stirred, cooled (-20° ) solution of 10.8 g. (0.026 mol.) of7-amino-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid in 220 ml. of 3% sodium bicarbonate and 220 ml. of acetone isdropwise added a solution of 3.66 g. (0.029 mol.) of methylthioacetylchloride in 52 ml. of acetone, during which time the pH of the reactionmixture is maintained at 8.0 by addition of 10% sodium hydroxide. Afteraddition the reaction mixture is stirred an additional 20 minutes at-15°, then is warmed to 25° and extracted with ether. The remainingaqueous phase is cooled, 250 ml. of ethyl acetate is added and theslurry is acidified with 3N hydrochloric acid. The layers are separatedand the aqueous phase is extracted twice more with ethyl acetate. Thecombined extracts are dried (MgSO₄) and evaporated to dryness to yieldthe title compound.

EXAMPLE 357-n-Propylthioacetamido-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

The title compound is prepared by substitution of n-propylthioacetylchloride in the procedure of Example 34 for methylthioacetyl chloride.

EXAMPLE 367-(α-Carboxy-2-thienylacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-5-cephem-4-carboxylicacid

7-Aminocephalosporanic acid (0.73 g., 2.68 mmol.) is suspended in 10 ml.of dry dimethylformamide and 1.12 ml. of triethylamine is added. Asolution of 0.91 g. (2.68 mmol.) of the activated ester ofα-t-butoxycarbonyl-2-thienylacetamide, prepared as described above, in 2ml. of dimethylformamide is added and the reaction mixture is stirred at25° for three hours. An additional 0.45 g. of activated ester is thenadded and the mixture is stirred another five hours. Ether (ca. 300 ml.)is added and the solution is decanted. The remaining material is washedwith ether, dissolved in water and the aqueous solution is extractedwith ethyl acetate. The pH of the ethyl acetate solution is adjusted to1.5 by addition of dilute hydrochloric acid. Extraction with ethylacetate and evaporation of the solvent gives7-(α-t-butoxycarbonyl-2-thienylacetamido)cephalosporanic acid.

A solution of 11.8 g. (0.06 mol.) of 1-sulfomethyltetrazole-5-thiol in120 ml. of water is added to a warm (45° ) solution of 19.9 g. (0.04mol.) of 7-(α-t-butoxycarbonyl-2-thienylacetamido)cephalosporanic acidin a mixture of 220 ml. of water and 8.4 g. (0.01 mol.) of sodiumbicarbonate. The temperature is raised to 65° and the pH maintained at7.4-7.6 by addition of aqueous sodium carbonate solution. After threehours, the reaction mixture is cooled to 10° and adjusted to pH 3.5 byaddition of dilute hydrochloric acid. The pH is adjusted to 7.0 byaddition of base and the aqueous solution is passed through a XAD-4resin column eluting with water and methanol to give the title compoundas its disodium salt.

7-(α-t-Butoxycarbonyl-2-thienylacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid disodium salt (1.0 g.) is stirred in a mixture of 10 ml. oftrifluoroacetic acid and 10 ml. of m-dimethoxybenzene at 25° for onehour. The mixture is evaporated to dryness and the residue is trituratedwith ether. The precipitated product is collected by filtration anddissolved in methanol. A sodium methoxide solution is added until pH7.0, the mixture is diluted with ether and the product is dissolved inwater and lyophilized to give7-(α-carboxy-2-thienylacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid trisodium salt.

The trisodium salt is converted to the title compound by treating thesalt with IR-120H ion exchange resin as previously described.

EXAMPLE 377-(2-Thienylacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

Reaction of 2.18 g. (0.01 mol.) of 1-sulfomethyltetrazole-5-thiol sodiumsalt, 5.23 g. (0.012 mol.) of 7-(2-thienylacetamido)cephalosporanic acidsodium salt and 0.84 g. (0.01 mol.) of sodium bicarbonate as describedin the procedure of Example 14 gave the title compound as thecorresponding disodium salt.

    C.sub.16 H.sub.14 N.sub.6 O.sub.7 S.sub.4 .2Na . 0.75 C.sub.2 H.sub.6 O

calculated: 34.81% C; 3.09% H; 13.56% N Found: 34.31% C; 3.03% H; 13.76%N

7-(2-Thienylacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid disodium salt is converted to the title compound as describedabove.

EXAMPLE 387-(2-Thienylacetamido)-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

Reaction of 5.68 g. (0.01 mol.) of 1-(2-sulfoethyl)tetrazole-5-thiolcyclohexylamine salt and 5.23 g. (0.0125 mol.) of7-(2-thienylacetamido)cephalosporanic acid sodium salt in 80 ml. ofwater containing sufficient sodium bicarbonate to maintain the pH at7.0, as described in the procedure of Example 14, gave the titlecompound.

EXAMPLE 397-Trifluoromethylthioacetamido-3-[1-(3-sulfamoylpropyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

Reaction of 1-(3-sulfamoylpropyl)tetrazole-5-thiol sodium salt, preparedas previously described, and7-trifluoromethylthioacetamidocephalosporanic acid sodium salt accordingto the procedure described in Example 14 gives the title compound as itssodium salt.

7-Trifluoromethylthioacetamido-3-[1-(3-sulfamoylpropyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid sodium salt is converted to the title compound as described inExample 14.

EXAMPLE 407-Amino-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

To a mixture of 97 g. (200 ml., 2.1 mol.) of formic acid, distilled fromanhydrous copper sulfate, and 37.5 ml. (0.4 mol.) of acetic anhydridewas added 25.0 g. (0.1 mol.) of 7-aminocephalosporanic acid. The mixturewas stirred at ambient temperature for 0.5 hour, then evaporated todryness. The residue was dissolved in ethyl acetate and the ethylacetate solution was filtered and evaporated to dryness to give aresidue which was recrystallized from ether-petroleum ether to give7-formamidocephalosporanic acid.

A mixture of 1.0 g. (3.3 mmol.) of 7-formamidocephalosporanic acid and0.7 g. (2.6 mmol.) of 1-sulfomethyltetrazole-5-thiol disodium salt in 15ml. of water was stirred at 65-70° for 3 hours while maintaining the pHat 7.0 by addition of sodium bicarbonate and/or hydrochloric acid. Themixture was cooled, acidified to pH 1.0 with hydrochloric acid andextracted with ethyl acetate. The extract was filtered and the filtratewas evaporated to dryness to give a residue which was dissolved in 30ml. of methanol. The methanol solution was filtered, 30-40 ml. ofisopropanol and ethanol were added, the solution was filtered again and100 ml. of ether was added. The precipitate was collected by filtrationand dried to give the title compound.

    C.sub.10 H.sub.12 O.sub.6 N.sub.6 S.sub.3 . 0.33 C.sub.3 H.sub.8 O . HCl . 2 H.sub.2 O

calculated: 27.0% C; 3.50% H; 16.95% N Found: 26.96% C; 3.23% H; 16.55%N

EXAMPLE 417-(4-Pyridylthioacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

(4-Pyridylthio)acetyl chloride (0.53 g., 2.8 mmol.) was dropwise addedto a mixture of 1.0 g. of7-amino-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid (mixture of sodium salt, free acid and sodium chloride) and 0.9 g.(9.0 mmol.) of triethylamine in 10 ml. of dry dimethylformamide. Thereaction mixture was stirred for 1.5 hour at -10°, then it was warmed toambient temperature and stirred for 1 hour. The mixture was filtered andthe filtrate was diluted with 200 ml. of ether-petroleum ether. Theprecipitate was collected by filtration and dissolved in warm methanol.The methanol solution was filtered and the filtrate was concentrated to10 ml. Ethanol (50 ml.) was added and the precipitate was collected byfiltration and washed with ether to give the title compound as thesodium salt. Additional amounts of the salt were obtained by addition ofether to the filtrate.

    C.sub.17 H.sub.16 N.sub.7 O.sub.7 S.sub.4 . Na . NaCl . C.sub.6 H.sub.15 N

calculated: 37.23% C; 4.21% H; 15.10% N Found: 36.48% C; 4.48% H; 15.43%N

7-(4-Pyridylthioacetmido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid sodium salt is converted to the title compound by proceduresdescribed above.

EXAMPLE 42

Acylation of7-amino-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid with an activated derivative of the following acids:

cyanoacetic acid

3-pyridylthioacetic acid

cyanomethylthioacetic acid

2,2,2-trifluoroethylsulfinylacetic acid

trifluoromethylsulfonylacetic acid

as described in the procedure of Example 41 gives the followingcompounds of this invention:

7-cyanoacetamido-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

7-(3-pyridylthioacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

7-cyanomethylthioacetamido-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

7-(2,2,2-trifluoroethylsulfinylacetmido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

7-trifluoromethylsulfonylacetamido-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid.

EXAMPLE 437-(D-α-Formyloxyphenylacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

7-Amino-3-(1-sulfomathyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid is reacted with the formate ester of D-mandeloyl chloride accordingto the procedure of Example 41 to give the title compound.

EXAMPLE 447-(D-α-Formyloxyphenylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

Substitution of an equivalent amount of7-amino-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid in the procedure of Example 43 for7-amino-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem4-carboxylicacid gives the title compound.

EXAMPLE 45

Using the procedures described above for the preparation of compounds ofthis invention, the following compounds were prepared:

7-(1-tetrazolylacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

    C.sub.13 H.sub.12 N.sub.10 O.sub.7 S.sub.3 . 2 Na

Calculated: 27.76% C; 2.15% H; 24.90% N Found: 27.55% C; 3.21% H; 23.00%N

7-(3-sydnoneacetamido)-3-(1-sulfomethyltetrazol-5ylthiomethyl)-3-cephem-4-carboxylicacid

    C.sub.14 H.sub.10 N.sub.8 O.sub.9 S.sub.3 . 2 Na . 3 H.sub.2 O . 2 CH.sub.4 O

calculated: 27.59% C; 3.72% H; 16.08% N Found: 27.59% C; 3.02% H; 15.67%N

7-trifluoromethylthioacetamido-3-[1-(4-sulfobutyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

    C.sub.16 H.sub.17 F.sub.3 N.sub.6 O.sub.7 S.sub.4 . 2 Na . H.sub.2 O

calculated: 29.36% C; 2.93% H; 12.84% N Found: 29.31% C; 3.33% H; 12.76%N

7-(2-thienylacetamido)-3-[1-(4-sulfobutyl)tetrazol5-ylthiomethyl]-3-cephem-4-carboxylicacid

    C.sub.19 H.sub.20 N.sub.6 O.sub.7 S.sub.4 . 2 Na . 2 H.sub.2 O . 2 C.sub.2 H.sub.6 O

calculated: 34.85% C; 3.69% H; 12.83% N; 19.59% S Found: 34.52% C; 3.88%H; 13.54% N; 19.09% S

7-(2-thienylacetamido-3-[1-(5-sulfopentyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid.

    C.sub.20 H.sub.22 N.sub.6 O.sub.7 S.sub.4 . 2 Na . 2.5 H.sub.2 O . 1.75 C.sub.2 H.sub.6 O

Calculated: 36.33% C; 4.77% H; 11.29% N; 17.23% S Found: 36.46% C; 4.30%H; 11.63% N; 17.30% S

EXAMPLE 467α-Methoxy-7β-(2-thienylacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

A solution of 1.28 g. (3 mmol.) of7α-methoxy-7β-(2-thienylacetamido)cephalosporanic acid sodium salt isdissolved in 50 ml. of water, 1.08 g. (4.5 mmol.) of1-sulfomethyltetrazole-5-thiol disodium salt is added and the solutionis heated at 70° until thin layer chromatography indicates consumptionof the starting material (ca. 5 hours). The reaction mixture ischromatographed on XAD-4 ion exchange resin with, after washing withwater, methanol as eluant. Evaporation of the methanol solution givesthe title compound as the disodium salt.

7α-Methoxy-7β-(2-thienylacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid disodium salt is converted to the title compound as describedabove.

EXAMPLE 477α-Methoxy-7β-trifluoromethylthioacetamido-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

To a cold solution of 5.25 g. (0.012 mol.) of7β-amino-7α-methoxycephalosporanic acid benzhydryl ester in 200 ml. ofmethylene chloride containing 1.79 g. (0.012 mol.) of N,N-diethylanilineis added dropwise over a 20 minute period a solution of 1.82 g. (0.012mol.) of trifluoromethylthioacetyl chloride in 50 ml. of methylenechloride. After stirring for 30 minutes, the mixture is extractedsuccessively with 5% aqueous sodium bicarbonate, 5% aqueous hydrochloricacid and finally with brine. The organic phase is dried (MgSO₄) and thesolvent evaporated to give7α-methoxy-7β-trifluoromethylthioacetamidocephalosporanic acidbenzhydryl ester.

7α-Methoxy-7β-trifluoromethylthioacetamidocephalosporanic acidbenzhydryl ester is dissolved in a cold mixture of trifluoroaceticacid-anisole (2:1) and the mixture is stirred for 1.5 hr. withoutexternal cooling. The solvent is evaporated in vacuo and the residualproduct is taken up in ethyl acetate, washed with water, dried (MgSO₄)and concentrated in vacuo to a small volume. This solution is addeddropwise to stirred petroleum ether to yield7α-methoxy7β-trifluoromethylthioacetamidocephalosporanic acid.

7α-Methoxy-7β-trifluoromethylthioacetamidocephalosporanic acid (2.2 g.,5 mmol.), is suspended in 75 ml. of water and 0.4 g. of solid sodiumbicarbonate is added until solution is complete. To this solution isadded 1.8 g. (7.5 mmol.) of 1-sulfomethyltetrazole-5-thiol disodium saltand the mixture is heated at 70° for 7 hours. The pH of the reactionmixture is maintained at 7.5 by dropwise addition of 3N hydrochloricacid as necessary. Progress of the reaction is monitored by thin layerchromatography and judged to be complete when tlc indicatesdisappearance of starting material. The reaction mixture ischromatographed on a column of XAD-4 resin and the product is elutedfrom the column with methanol. Evaporation of the methanol solution give7α-methoxy-7β-trifluoromethylthioacetamido-3(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid disodium salt.

The disodium salt is converted to the title compound by proceduresdescribed hereinabove.

EXAMPLE 487β-D-Mandelamido-7α-methoxy-3-(1-sufomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

A cold solution of 2.6 g. (6 mmol.) of7β-amino-7α-methoxycephalosporanic acid benzhydryl ester in 100 ml. ofmethylene chloride containing 0.9 g. (6 mmol.) of N,N-diethylaniline istreated dropwise over a 15 minute period with a solution of 1.7 g. (6mmol.) of D-O-dichloroacetylmandeloyl chloride in 25 ml. of methylenechloride. The reaction mixture is allowed to come to room temperaturewith stirring and then is extracted successively with 5% aqueous sodiumbicarbonate, 5% hydrochloric acid and brine. The organic phase is driedand evaporated in vacuo. The residue is dissolved in coldtrifluoroacetic acid-anisole (2:1) and the mixture is sitrred at ambienttemperature for 1 hour. The mixture is evaporated in vacuo and theresidue is dissolved in 5% aqueous sodium bicarbonate. The pH is raisedto 9-9.3 by addition of 5% aqueous sodium carbonate and maintained therefor 30 minutes to complete cleavage of the dichloroacetyl group. Thesolution is cooled in ice, layered with ethyl acetate and acidified topH 2.0 with dilute hydrochloric acid. The layers are separated and aftera second extraction of the aqueous layer with ethyl acetate the organicphases are combined, dried and evaporated in vacuo to yield7β-D-mandelamido-7α-methoxycephalosporanic acid.

7β-D-mandelamido-7α-methoxycephalosporanic acid (2.2 g., 5 mmol.) issuspended in 75 ml. of water and solid sodium bicarbonate is added untilall of the acid has dissolved. To this is added 1.8 g. (7.5 mmol.) of1-sulfomethyltetrazole-5-thio disodium salt and the mixture is heated at70° for 7 hours. The pH of the reaction mixture is maintained at 7.5 byaddition of 3N hydrochloric acid. Chromatography of this solution onXAD-4 resin while eluting with methanol gives, upon evaporation of themethanol, the title compound as its disodium salt.

7β-D-Mandelamido-7α-methoxy-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid disodium salt is converted to the title compound by proceduresdescribed hereinabove.

EXAMPLE 497α-Methoxy-7β-(D-α-aminophenylacetamido)-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

To a solution of 5.3 g. (0.012 mol.) of7β-amino7α-methoxycephalosporanic acid p-nitrobenzyl ester in 200 ml. ofmethylene chloride is added 3.0 g. (0.012 mol.) ofD-α-t-butoxycarbonylaminophenylacetic acid and 2.5 g. (0.012 mol.) ofdicyclohexylcarbodiimide. The mixture is stirred for 18 hours at ambienttemperature then filtered. The filtrate is evaporation in vacuo and theresidue is dissolved in methanoltetrahydrofuran and hydrogenated over 5%palladium on carbon to give7β-(D-α-t-butoxycarbonylaminophenylacetamido)-7α-methoxycephalosporanicacid.

7β-(D-α-t-Butoxycarbonylaminophenylacetamido)7α-methoxycephalosporanicacid (2.68 g., 5 mmol.) is dissolved in 75 ml. of water by adding 0.4 g.of solid sodium bicarbonate. 1-(2-Sulfoethyl)tetrazole-5-thiol disodiumsalt (1.9 g., 7.5 mmol.) is added and the reaction mixture is heated at70° until thin layer chromatography indicates that the starting materialhas disappeared. The reaction mixture is chromatographed on XAD-4resinand eluted with methanol. Evaporation of the methanol solution gives7α-methoxy-7β-(D-α-t-butoxycarbonylaminophenylacetamido)-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid disodium salt.

The disodium salt is suspended in 1:1 trifluoroacetic acid-ansiole andstirred at ambient temperature for two hours. Excess trifluoroaceticacid is removed by evaporation, the residue is triturated with ether andthe resulting precipitate is collected by filtration and stirred withacetonitrile to give the title compound as its trifluoroacetic acidsalt.

An aqueous solution of the trifluoroacetic acid salt is brought to pH 7by addition of 5% aqeuous sodium bicarbonate then chromatographed onXAD-4 resin with methanol as eluant. The solid material obtained afterevaporation of the methanol is dissolved in water and the aqueoussolution is passed through a cation exchange column (IR-120H).Lyophilization of the eluted material gives the title compound.

EXAMPLE 507β-D-Mandelamido-7α-methoxy-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid

To a suspension of 21.1 g. (0.05 mol.) of7-amino3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4carboxylicacid in 500 ml. of dry methylene chloride is added in one portion asolution of 30.0 g. (0.15 mol.) of O-t-butyldiisopropylpseudourea in 50ml. of methylene chloride. The mixture is stirred at ambient temperaturefor 24 hours. The precipitate is removed by filtration and the filtrateevaporated to a residue which is taken up in 200 ml. of benzene andfiltered again. The filtrate is extracted with three 100 ml. portions ofcold 1N hydrochloric acid. The acidic aqueous extracts are layered withethyl acetate and the pH adjusted to 7.5 with solid sodium bicarbonate.The organic layer is separated and the aqueous phase is extracted withtwo 150 ml. portions of ethyl acetate. The combined extracts are dried(MgSO₄), filtered and evaporated to give7-amino-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid t-butyl ester.

A solution of 1.91 g. (4 mmol.) of7-amino-3[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid t-butyl ester and 0.94 g. (4 mmol.) of3,5-di-t-butyl-4-hydroxybenzaldehyde in 150 ml. of dry benzene isrefluxed for ca. 4 hours under a Dean-Stark trap until no more waterseparates. The solution is evaporated under reduced pressure to give aresidue which is dissolved in 150 ml. of 1,2-dichloroethane and cooledto 0-5° in an ice bath. Freshly prepared lead dioxide (5 g.) is added in1 g. portions over 30 minutes and the reaction is stirred in the colduntil complete consumption of the starting material is indicated by thinlayer chromatography. The mixture is then filtered through Celite andthe filter cake is washed with two 30 ml. portions of cold1,2-dichloroethane. The filtrate is treated with 30 ml. of dry methanol(distilled from magnesium) and the reaction is stirred at ambienttemperature for 3 hours until complete consumption of the intermediateand formation of a new product is shown by thin layer chromotography.The reaction mixture is evaporated to dryness and the residue is takenup in 50 ml. of methanol and treated with 4.0 g. of Girard reagent T(trimethylaminoacetohydrazide chloride). This solution is stirred atambient temperature for 3 hours and evaporated under vacuum to give asolid residue which is partitioned between 150 ml. of ethyl acetate and100 ml. of 20% aqueous sodium chloride solution. The organic phase iswashed with three 100 ml. portions of 10% aqueous sodium chloride, two100 ml. portions of water and 100 ml. of a saturated sodium chloridesolution. The organic phase is dried (MgSO₄), filtered and evaporated todryness to give 7β-amino-7α-l-methoxy-3-[1-(2-sulfamoyethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid t-butyl ester.

A solution of 0.95 g. (2 mmol.) of7β-amino-7α-methoxy-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid t-butyl ester and 0.30 g. (2 mmol.) of N,N-diethylaniline in 100ml. of dry methylene chloride is stirred at 0 -5° while 0.56 g. (2mmol.) of D-O-dichloroacetylmandeloyl chloride in 10 ml. of methylenechloride is added dropwise over a 10 minute period. The mixture isstirred in the cold for 30 minutes then warmed to room temperature andstirred for an additional 30 minutes. The solution is washed with 50 ml.of cold dilute hydrochloric acid and 50 ml. of cold 5% aqueous sodiumbicarbonate, dried (MgSO₄) and evaporated to dryness. The residue isdissolved in a mixture of 10 ml. of trifluoroacetic acid and 2 ml. ofm-dimethoxybenzene and stirred at ambient temperature for 2 hours. Theexcess trifluoroacetic acid is evaporated under vacuum and the residueis partitioned between 50 ml. of ether and 50 ml. of water. The pH isadjusted to 9.3-9.5 with 5% aqueous sodium carbonate and the organicphase is separated and discarded. The aqueos phase is stirred at pH9.3-9.5 for 30 minutes, extracted with 50 ml. of ethyl acetate which isdiscarded, layered with fresh ethyl acetate and adjusted to pH 1.5 withdilute hydrochloric acid. The aqueous layer is extracted with three 50ml. portions of ethyl acetate and the combined extracts are dried andevaporated to a small volume. Petroleum ether is added dropwise toprecipitate the title compound which is collected and dried.

EXAMPLE 51

An injectable pharmaceutical composition is formed by adding sterilewater or sterile saline solution (2 ml.) to 500 mg. of7-D-mandelamido-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid sodium salt.

Pharmaceutical compositions of the other antibacterial compoundsdisclosed above may be formulated in a similar manner.

What is claimed is:
 1. A compound of the formula: ##STR9## in which:each individual R¹ hydrogen or lower alkyl;n is one to ten; and R² ishydroxy, amino, lower alkylamino or di(lower)alkylamino, or a saltthereof.
 2. A compound according to claim 1 in which R¹ is hydrogen andn is one to five.
 3. A compound according to claim 2 in which R² ishydroxy.
 4. A compound according to claim 2 in which R² is amino.
 5. Acompound according to claim 2, said compound being1-(2-N-t-butylsulfamoylethyl)tetrazole-5-thiol.
 6. A compound accordingto claim 3, said compound being 1-sulfomethyltetrazole-5-thiol.
 7. Acompound according to claim 3, said compound being1-(2-sulfoethyl)tetrazole-5-thiol.
 8. A compound according to claim 3,said compound being 1-(5-sulfopentyl)tetrazole-5-thiol.
 9. A compoundaccording to claim 4, said compound being1-(2-sulfamoylethyl)tetrazole-5-thiol.
 10. A compound according to claim4, said compound being 1-(5-sulfamoylpentyl)tetrazole-5-thiol.
 11. Thesodium salt of 1-sulfomethyltetrazole-5-thiol.